DNA-Based Biomarkers for Multiple Sclerosis

描述（由申请人提供）：多发性硬化症（MS）是一种慢性炎症性神经退行性疾病，影响中枢神经系统，可能导致10-15年内严重残疾。MS通过临床症状和通过磁共振成像检测的炎症的存在来诊断。其他疾病也有类似的症状，因此MS的诊断具有挑战性。缺乏用于诊断、监测或预测结果的明确和客观的生物标志物使MS检测和MS患者的治疗复杂化。我们建议使用我们实验室开发的一种新的检测方法来分析DNA甲基化，以确定每个标本中多个基因中的MS特异性DNA甲基化变化，并创建一个复合生物标志物，将基因与最多样化的甲基化模式相结合。MS患者的血脑屏障被破坏，来自死亡细胞的基因组DNA被释放到血流中，因此该测定可以基于无细胞循环DNA。本项目的具体目的是：（1）建立一种基于无细胞循环血浆DNA甲基化谱的复合生物标志物，用于检测缓解期复发缓解型MS患者;（2）基于无细胞循环血浆DNA的DNA甲基化谱建立用于检测复发-缓解型MS患者在恶化期间的复合生物标志物，并将其与目标1中建立的生物标志物进行比较;（3）比较复发-缓解型MS和继发-进展型MS患者的DNA甲基化谱，并确定对进展特异的生物标志物。该测定允许使用来自0.15 ml血浆的DNA同时检测56个基因启动子中的甲基化。该测定成功地检测到MS患者血浆中DNA的MS特异性变化。该项目的完成将产生一套新的客观的非依赖性生物标志物，其基于快速，简单和廉价的程序，并利用简单，安全和微创的标本采集。目前还没有关于MS特异性DNA甲基化的报道，因此该项目将是第一个解决这一缺陷的项目。无细胞循环DNA用于MS检测的实用性从未报道过，因此该项目将是第一个利用这一机会的项目。因此，该项目的其他组成部分是没有先例的：将首次开发不同形式RR-MS的生物标志物，预测发作的生物标志物和疾病进展为SP-MS的生物标志物。多发性硬化症（MS）是一种中枢神经系统慢性炎症性疾病，可导致严重残疾和死亡。MS的诊断是困难的，因为疾病的初始阶段没有明显的症状，甚至根本没有任何症状，也没有MS特异性的客观标志物。在这个项目中，我们将使用我们实验室开发的一种新技术来评估MS患者血液中的DNA修饰（甲基化），活动性MS（发作或加重）和稳定的MS（缓解）。完成的项目将提供第一个基于MS特异性DNA甲基化的客观生物标志物，可用于MS的诊断，治疗成功的评估和结果的预测。

DESCRIPTION (provided by applicant): Multiple sclerosis (MS) is a chronic, inflammatory neurodegenerative disease that affects the central nervous system and may lead to severe disability within 10-15 years. MS is diagnosed by clinical symptoms and presence of inflammation detected by magnetic resonance imaging. Other diseases present with similar symptoms, so diagnosis of MS is challenging. The lack of clear and objective biomarkers for diagnosis, monitoring, or prediction of outcomes complicates MS detection and treatment of MS patients. We propose to use a novel assay developed in our laboratory for analysis of DNA methylation to identify MS-specific changes of DNA methylation in multiple genes within each specimen, and to create a composite biomarker combining genes with the most diverse patterns of methylation. The blood-brain barrier is disrupted in MS patients, genomic DNA from dead cells is released into the bloodstream, so the assay can be based on cell-free circulating DNA. Specific Aims of the project are: (1) to establish a composite biomarker for detection of patients with relapsing-remitting MS in remission based on DNA methylation profile of cell-free circulating plasma DNA; (2) to establish a composite biomarker for detection of patients with relapsing-remitting MS during exacerbations based on DNA methylation profile of cell-free circulating plasma DNA, and to compare it with the biomarker established in Aim 1; (3) to compare DNA methylation profiles of patients with relapsing- remitting MS and secondary-progressive MS and to determine biomarkers specific for progression. The assay allows simultaneous detection of methylation in 56 gene promoters using DNA from 0.15 ml of plasma. The assay successfully detected MS-specific changes in DNA from plasma of MS patients. Completion of this project will produce a novel set of objective observer-independent biomarkers based on a fast, simple, and inexpensive procedure, and utilizing a simple, safe, and minimally invasive specimen collection. There are no reports on MS-specific DNA methylation, so this project will be the first to address this deficiency. The utility of cell-free circulating DNA for testing in MS has never been reported, so this project will be the first to use this opportunity. Consequently, other components of this project are without precedent: a biomarker for different forms of RR-MS, a biomarker for prediction of an attack, and a biomarker for disease progression to SP-MS will be developed for the first time. Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system, which can lead to severe disability and death. Diagnosis of MS is difficult because initial stages of the disease do not have obvious symptoms or even any symptoms at all, and there are no objective markers specific for MS. In this project we will use a new technique developed in our laboratory to evaluate DNA modifications (methylation) in blood of MS patients with active MS (attack or exacerbation) and stable MS (remission). Completed project will provide the first objective biomarker based on MS-specific DNA methylation, which can be used for diagnosis of MS, for evaluation of treatments success, and for prediction of outcomes.